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. 2017 Aug 30;45(18):10751–10763. doi: 10.1093/nar/gkx744

Figure 6.

Figure 6.

AtPolIs are low-fidelity DNAPs that bypass an AP site following the A rule. (A) Nucleotide insertion opposite a canonical template and an AP site. (A) Single nucleotide primer extension reactions using a template containing a canonical (dGMP) (lanes 1–5) or an AP site (lanes 6–10) following the 3′-OH end of the primer. In the reactions incubated with AtPolIB in the presence of dATP, dTTP and dGTP opposite a template G a band corresponding to nucleotide addition is synthetized (lanes 1–5). In the presence of dGTP, this band can be further extended (lane 5). In the reaction opposite a non-instructional AP site, dAMP is preferentially incorporated. dGMP and dTTP are incorporated with similar efficiencies, but dCTP is not efficiently used as a substrate (lanes 6–10). (B) AtPolIB is a low fidelity enzyme. Nucleotide incorporation by AtPolIB opposite all template bases. The identity of the template base is indicated by an X and the incorporated dNMP (A, T, G, C) is indicated. In all substrates, erroneous base incorporation and extension is observed. Incoming dNTPs were present at 100 μM.